WO2017215428A1 - Procédé de préparation d'une plaque de matrice de micro-canaux, dispositif d'obtention de gouttes de liquide à l'aide de la plaque de matrice de micro-canaux, et procédé de génération de gouttes de liquide - Google Patents

Procédé de préparation d'une plaque de matrice de micro-canaux, dispositif d'obtention de gouttes de liquide à l'aide de la plaque de matrice de micro-canaux, et procédé de génération de gouttes de liquide Download PDF

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Publication number
WO2017215428A1
WO2017215428A1 PCT/CN2017/085891 CN2017085891W WO2017215428A1 WO 2017215428 A1 WO2017215428 A1 WO 2017215428A1 CN 2017085891 W CN2017085891 W CN 2017085891W WO 2017215428 A1 WO2017215428 A1 WO 2017215428A1
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WO
WIPO (PCT)
Prior art keywords
array plate
microchannel array
liquid
hole
oil
Prior art date
Application number
PCT/CN2017/085891
Other languages
English (en)
Chinese (zh)
Inventor
黄岩谊
陈子天
廖珮宇
张芳丽
Original Assignee
北京大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 北京大学 filed Critical 北京大学
Priority to US16/309,092 priority Critical patent/US20190358625A1/en
Publication of WO2017215428A1 publication Critical patent/WO2017215428A1/fr
Priority to US17/856,749 priority patent/US20220339620A1/en

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/02—Burettes; Pipettes
    • B01L3/0241—Drop counters; Drop formers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F23/00—Mixing according to the phases to be mixed, e.g. dispersing or emulsifying
    • B01F23/40—Mixing liquids with liquids; Emulsifying
    • B01F23/41—Emulsifying
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F23/00—Mixing according to the phases to be mixed, e.g. dispersing or emulsifying
    • B01F23/40—Mixing liquids with liquids; Emulsifying
    • B01F23/41—Emulsifying
    • B01F23/414—Emulsifying characterised by the internal structure of the emulsion
    • B01F23/4145—Emulsions of oils, e.g. fuel, and water
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F25/00—Flow mixers; Mixers for falling materials, e.g. solid particles
    • B01F25/20—Jet mixers, i.e. mixers using high-speed fluid streams
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F35/00—Accessories for mixers; Auxiliary operations or auxiliary devices; Parts or details of general application
    • B01F35/71—Feed mechanisms
    • B01F35/717—Feed mechanisms characterised by the means for feeding the components to the mixer
    • B01F35/71725—Feed mechanisms characterised by the means for feeding the components to the mixer using centrifugal forces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/061—Counting droplets
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0673—Handling of plugs of fluid surrounded by immiscible fluid
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/12—Specific details about manufacturing devices
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0829—Multi-well plates; Microtitration plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0832—Geometry, shape and general structure cylindrical, tube shaped
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/16—Surface properties and coatings
    • B01L2300/161—Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/16—Surface properties and coatings
    • B01L2300/161—Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
    • B01L2300/165—Specific details about hydrophobic, oleophobic surfaces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0409—Moving fluids with specific forces or mechanical means specific forces centrifugal forces

Definitions

  • the etching the microchannel array plate blank with the etching solution to obtain a microchannel array plate with a through hole includes: the microchannel array plate blank is Ultrasonic soaking in the nitric acid solution for a certain period of time, taking out the microchannel array plate blank, washing and soaking in the caustic soda solution for a certain time, then continuing to corrode in the acid solution, alternately reciprocating; the etching solution is doped with a small amount of fluoride ions.
  • the hydrophobic treatment comprises: modifying the surface of the glass by at least one of a method such as chemical vapor deposition, immersion, and solvent evaporation.
  • the first liquid is an aqueous phase liquid, which is a sample for biological reaction, including a mixture for a digital chain enzymatic reaction, a cell suspension , bacterial suspension, DNA solution for genomic amplification, mixture for RNA reverse transcription, mixture for protein crystallization, mixture for inorganic salt crystallization, pathogen solution or suspension, for A mixed solution of a polymerization reaction, a mixed solution for a gelation reaction, etc.
  • the second liquid is a surface active liquid phase liquid.
  • the oil phase in the second liquid is mineral oil (such as low boiling mineral oil, light mineral oil, etc.), silicone oil (such as oligodimethyl Siloxane, cyclopentasiloxane, aliphatic siloxane, phenyl siloxane, fluorosilicone, etc.), fatty acid glyceride (glycerol dilaurate, glyceryl oleate, glyceryl linoleate) , glyceryl stearate, glycerol linolenate, glyceryl isostearate, glyceryl sorbate, etc.), carbonic acid diester (such as bis-methyl-octyl carbonate, dihexadecyl carbonate, double sorbate) Alcohol ester, bis 2-ethylhexyl carbonate, bis 2-ethyloctyl carbonate, bis 2-ethyl
  • the surfactant in the second liquid is sodium cetylsulfonate, 20, twenty one, 40, 60, 61, 65, 80, 20, 40, 60, 80, 83, 85, 120, We09, Em90, Em120, Em180, Dow 5200, Dow ES-5300, Dow em ⁇ Lsifier 10, SML, WO 7, GI 34, GI PDI, One or more of the Alkanol S 2Pellets are mixed.
  • the collecting device is a centrifuge tube.
  • the connecting member includes a blind hole formed from the upper end surface, the blind hole forming an inner end surface in the connecting member, and the inner wall of the blind hole has
  • the internal thread is a female thread
  • the diameter of the blind hole is adapted to the outer diameter of the microchannel array plate, and the inner end surface to the lower end surface form a through hole
  • the through hole is a tapered hole whose diameter gradually increases from the top to the bottom.
  • the tapered hole has a minimum diameter smaller than an inner diameter of the inner end surface, and in use, the microchannel array plate is located between a lower end surface of the bolt and the inner end surface of the connecting member.
  • the inside of the bolt is a through hole
  • the through hole is a large diameter circular hole, a tapered hole, and a small diameter circular hole from top to bottom.
  • the tapered hole connects the large diameter circular hole and the small diameter circular hole, and the tapered hole has an angle of 30 to 50 degrees.
  • Fig. 6 is a bright field photograph and a fluorescent photograph after 30 rounds of PCR were carried out by using a microplate to generate droplets of the A sample.
  • Embodiments of the present invention provide a method of preparing a microchannel array panel.
  • the split glass rod is fused to one another at 800 degrees Celsius to 3000 degrees Celsius under the condition of external force traction at both ends, and then becomes a thin glass rod or filament under external force traction.
  • the secondary drawing is repeated the above drawing step, and the drawn glass rods or filaments are arranged again and then drawn.
  • the glass rods of the appropriate diameter of the drawn glass rods are cut and polished to obtain the glass sheets, which can be used as raw materials for the microchannel array sheets.
  • the thinned glass rod was cut into small pieces of about 1 mm, and then polished several times to obtain a microchannel array plate blank.
  • the surface of the glass has some silicon-oxygen bonds and silanols, which are hydrophilic. Due to the presence of these hydrophilic groups, water and aqueous solutions are spread over the surface of the glass to provide a certain contact area with the surface of the glass. In order to make the water from the microchannel have a spherical surface for smooth detachment, and at the same time make the droplet size of each detachment uniform, it is necessary to exclude the influence of water or aqueous solution and glass surface infiltration, which requires microchannel array. The plate is subjected to a hydrophobic treatment.
  • fluorosilanes are used in small amounts and can meet the hydrophobic properties in a small amount.
  • the hydrophobic surface of the microchannel array plate is one of the keys to the regular suspension breakage of the droplets.
  • the glass surface is typically modified by chemical vapor deposition.
  • the steps of the modification generally include the steps of washing with concentrated sulfuric acid hydrogen peroxide, washing with ultrapure water, drying, surface activation of oxygen radicals, chemical vapor fumigation, aging and ultrapure water cleaning.
  • gas phase fumigation it can also be immersed, and the steps thereof generally include washing, oxygen radical activation, soaking, aging, post-cleaning, and the like.
  • microemulsion droplets For microporous plates with a pore size of 6 ⁇ m, different sizes of microemulsion droplets can be obtained by centrifugal force of different sizes.
  • the bolts and connecting parts can be made of PEEK.
  • sealing gasket between the microchannel array plate and the inner end surface of the connecting member, which is a ring having an outer diameter of about 5 mm and an inner diameter of about 3 mm.
  • the thickness is from 0.2 mm to 2 mm, for example 1 mm. It can be obtained by mechanical finishing from PEEK (polyetheretherketone) plastic without glass fiber, or it can be cut with soft board. Among them, the gasket made of Teflon has the best effect.
  • the material of the gasket can also be rubber.
  • the microchannel microchannels of the microchannel microchannel array plate can be arbitrarily designed with different diameters.
  • the size of the droplet generation can be finely adjusted by centrifugal force.
  • the droplet size is adjusted when the droplet is generated by centrifugation on the microchannel microchannel array plate.
  • the assembly drawing is shown in Figure 5.
  • the positions of the bolts, washers, microchannel array plates and connecting members are the same as in Figs. 4 and 6 are 200 ⁇ L centrifuge tubes, 7 is a conversion bracket, and 8 is a 1.5 mL centrifuge tube.
  • the conversion bracket is 3D printed.

Abstract

L'invention porte également sur un procédé de préparation d'une plaque de matrice de micro-canaux, comprenant : (1) l'agencement d'une première tige de verre de fibre optique et d'une seconde tige de verre de fibre optique étroitement, et à faire fondre les deux tiges de verre à une température élevée pour obtenir une tige de verre fondue, à étirer la tige de verre fondue au moins une fois en une tige de verre plus longue et plus fine que la tige de verre fondue, et à découper la tige de verre étirée de manière fine en petits morceaux afin d'obtenir un blanc de plaque de matrice de micro-canaux, où la résistance à la corrosion de la première tige de verre de fibre optique et de la seconde tige de verre de fibre optique au même liquide corrosif étant différente; (2) à corroder le blanc de plaque de matrice de micro-canaux à l'aide d'un liquide corrosif pour obtenir un produit brut de plaque de matrice de micro-canaux avec des trous traversants; et (3) à effectuer un traitement hydrophobe sur le produit brut de plaque de matrice de micro-canaux afin d'obtenir la plaque de matrice de micro-canaux.
PCT/CN2017/085891 2016-06-12 2017-05-25 Procédé de préparation d'une plaque de matrice de micro-canaux, dispositif d'obtention de gouttes de liquide à l'aide de la plaque de matrice de micro-canaux, et procédé de génération de gouttes de liquide WO2017215428A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US16/309,092 US20190358625A1 (en) 2016-06-12 2017-05-25 Method for preparing micro-channel array plate, device for obtaining liquid drops using the micro-channel array plate, and method for generating liquid drops
US17/856,749 US20220339620A1 (en) 2016-06-12 2022-07-01 Method for preparing micro-channel array plate, device for obtaining liquid drops using the micro-channel array plate, and method for generating liquid drops

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201610409019.0A CN106076443B (zh) 2016-06-12 2016-06-12 一种微通道阵列板的制备方法、用其来获得液滴的装置和液滴产生方法
CN201610409019.0 2016-06-12

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US16/309,092 A-371-Of-International US20190358625A1 (en) 2016-06-12 2017-05-25 Method for preparing micro-channel array plate, device for obtaining liquid drops using the micro-channel array plate, and method for generating liquid drops
US17/856,749 Continuation US20220339620A1 (en) 2016-06-12 2022-07-01 Method for preparing micro-channel array plate, device for obtaining liquid drops using the micro-channel array plate, and method for generating liquid drops

Publications (1)

Publication Number Publication Date
WO2017215428A1 true WO2017215428A1 (fr) 2017-12-21

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Country Link
US (2) US20190358625A1 (fr)
CN (1) CN106076443B (fr)
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3837345A4 (fr) * 2018-10-01 2022-05-18 Molarray Research Inc. Pointe de micro-pipette pour former des micro-gouttelettes
US11814619B2 (en) 2021-06-04 2023-11-14 Enumerix, Inc. Compositions, methods, and systems for single cell barcoding and sequencing
US11834714B2 (en) 2021-12-20 2023-12-05 Enumerix, Inc. Detection and digital quantitation of multiple targets

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CN106076443B (zh) * 2016-06-12 2019-05-24 北京天天极因科技有限公司 一种微通道阵列板的制备方法、用其来获得液滴的装置和液滴产生方法
CN110505918B (zh) * 2017-02-13 2022-06-14 生物辐射实验室股份有限公司 用于形成乳状液阵列的系统、方法以及装置
CN109060736A (zh) * 2018-06-27 2018-12-21 北京天天极因科技有限公司 用于透明化液滴成像的光片荧光显微成像装置及检测方法
CN108675646B (zh) * 2018-07-23 2021-08-31 北方夜视技术股份有限公司 具有大粗糙度通道内表面的微孔光学元件及其制备方法
US11242558B2 (en) 2020-04-15 2022-02-08 Enumerix, Inc. Systems and methods for generation of emulsions with suitable clarity with applications of use
CN111545260B (zh) * 2020-05-10 2022-05-24 天津德祥生物技术有限公司 处理微通道内表面的方法
CN111863582B (zh) * 2020-07-24 2022-04-22 北方夜视技术股份有限公司 超声悬浮旋转式微通道板腐蚀方法
CN112619719B (zh) * 2020-12-04 2022-03-29 深圳先进技术研究院 用于数字pcr的液滴生成微型装置
CN114058495A (zh) * 2021-10-29 2022-02-18 浙江大学 一种工业级非对称液滴发生装置及数字核酸扩增检测系统

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3837345A4 (fr) * 2018-10-01 2022-05-18 Molarray Research Inc. Pointe de micro-pipette pour former des micro-gouttelettes
US11814619B2 (en) 2021-06-04 2023-11-14 Enumerix, Inc. Compositions, methods, and systems for single cell barcoding and sequencing
US11834714B2 (en) 2021-12-20 2023-12-05 Enumerix, Inc. Detection and digital quantitation of multiple targets

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US20220339620A1 (en) 2022-10-27
CN106076443A (zh) 2016-11-09
US20190358625A1 (en) 2019-11-28
CN106076443B (zh) 2019-05-24

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